Bis-hydrazones of daunomycin and adriamycin

ABSTRACT

Bis-hydrazones of daunomycin and adriamycin having useful antitumor characteristics and the structure ##STR1## WHEREIN N IS AN INTEGER HAVING A VALUE OF FROM 0 THROUGH 8 AND THE RADICALS INDICATED BY R and R&#39; each represent hydrogen except that in those instances in which n has a value of 2, the R groups, if not hydrogen, may be methyl radicals (with R&#39; remaining as hydrogen), and the R&#39; groups, if not hydrogen, may be hydroxy radicals (with R remaining as hydrogen).

The invention described herein was made in the course of or under acontract with the U.S. Department of Health, Education, and Welfare.

BACKGROUND OF INVENTION

The structures of the present invention represent a new class of doublyintercalating derivatives of daunomycin and adriamycin. Recent studiesfrom several laboratories [especially that by Canellakis et al.,Biophysica Acta, 418, 277-314 (1976)] have demonstrated that this modeof DNA binding is associated with important biological properties(especially cytotoxic and antitumor properties of a series ofbis-9-acridyl compounds). Use of the helical DNA-adriamycin molecularmodel, which has proven useful in the past in this project, suggestedthat bis-derivatives of the type disclosed could form a unique doublyintercalated complex.

SUMMARY OF INVENTION

The present invention rests on discovery of novel compounds havinguseful antitumor characteristics which possess the following generalstructure ##STR2## wherein n is an integer having a value of from 0through 8 and the radicals indicated by R and R' each represent hydrogenexcept that in those instances in which n has a value of 2, the Rgroups, if not hydrogen, may be methyl radicals (with R' remaining ashydrogen), and the R' groups, if not hydrogen, may be hydroxy radicals(with R remaining as hydrogen), together with their pharmaceuticallyacceptable acid addition salts.

    __________________________________________________________________________    Representative compounds which fall within the scope of the present           invention include:                                                            __________________________________________________________________________    R═R'═H,n=2                                                                      Bis(daunomycin) Succinylhydrazone Dihydrochloride (I)               R═R'═H,n=3                                                                      Bis(daunomycin) Glutarylhydrazone Dihydrochloride (II)              R═R'═H,n=4                                                                      Bis(daunomycin) Adipylhydrazone Dihydrochloride (III)               R═R'═H,n=6                                                                      Bis(daunomycin) Suberylhydrazone Dihydrochloride (IV)               R═R'═H,n=8                                                                      Bis(daunomycin) Sebacylhydrazone Dihydrochloride (V)                R═Me, R'═H,n=2                                                                  Bis(N,N-dimethyldaunomycin) Succinylhydrazone Dihydrochloride                 (VI)                                                                R═H,R'═OH,n=2                                                                   Bis(adriamycin) Succinylhydrazone Dihydrochloride (VII)             R═R'═H,n=1                                                                      Bis(daunomycin) Malonylhydrazone Dihydrochloride (VIII)             R═R'═H,n=0                                                                      Bis(daunomycin) Oxalylhydrazone Dihydrochloride (IX)                R═R'═H,n=5                                                                      Bis(daunomycin) Pimelylhydrazone Dihydrochloride                    __________________________________________________________________________              (X)                                                             

Compounds (I) through (X) enumerated above form the subject ofcorresponding examples 1 through 10, said examples including the detailsof the preparation of each compound as well as its characteristics. Itwill be noted that these compounds are prepared in the form of an acidaddition salt with the free NH₂ -or N(CH₃)₂ - groups of the compounds.These acid addition salts (prepared as those of HCl) are preferably thepharmaceutically acceptable, nontoxic addition salts with suitableacids, such as those with inorganic acids, for example, hydrochloric,hydrobromic, nitric, sulphuric and phosphoric acids, and with organicacids, such as organic carboxylic acids, for example, glycolic, maleic,hydroxymaleic, malic, tartaric, citric, salicyclic acids, and organicsulphonic acids, for example, methanesulphonic and toluene-p-sulphonicacids.

The compounds are preferably employed in the salt form since they haveadequate solubility in water. However, they can be employed in thenon-acid condition.

An acid addition salt can be converted into the free compound accordingto known methods, for example, by treating it with a base, such as witha metal hydroxide or alkoxide, for example, an alkali metal or alkalineearth metal hydroxide, for example, lithium hydroxide, sodium hydroxide,potassium hydroxide or calcium hydroxide; with a metal carbonate, suchas an alkali metal or an alkaline earth metal carbonate or hydrogencarbonate, for example sodium, potassium or calcium carbonate orhydrogen carbonate; with ammonia; or with a hydroxyl ion exchange resin,or with any other suitable reagent.

An acid addition salt may also be converted into another acid additionsalt according to known methods; for example, a salt with an inorganicacid may be treated with a metal salt, for example a sodium, barium orsilver salt, of an acid in a suitable diluent, in which a resultinginorganic salt is insoluble and is thus removed from the reactionmedium. An acid addition salt may also be converted into another acidaddition salt by treatment with an anion exchange preparation.

In the treatment of cancer, the compounds of this invention or the saltsthereof can be administered by any available route, including oral andparenteral (intravenous, intraperitoneal, subcutaneous, andintramuscular) administration. The amounts administered are thoseefficient to ameliorate the cancer.

Apart from requiring an extended reaction time, the compounds of thisinvention can readily be prepared by reacting the adriamycin ordaunomycin starting materials (in suitable salt form) with theappropriate dihydrazide reactant, the reaction proceeding in anappropriate solvent (e.g., methanol) at room temperatures. Thus, when nis to have a value of 2 or 3, for example, the reactants employed aresuccindihydrazide and glutardihydrazide, respectively.

EXAMPLE 1 Bis(daunomycin) Succinylhydrazone Dihydrochloride (I)

A mixture of 1.13 g (2.0 mmol) of daunomycin hydrochloride (XII) and0.146 g (1.0 mmol) of succinic acid dihydrazide in 120 ml of methanolwas stirred at room temperature in the dark for three days. The reactionmixture was concentrated in vacuo to about 50 ml and then stirred atroom temperature for two additional days. To the stirred solution wasadded 100 ml of ether dropwise; the resulting precipitate was collected,washed with ether and dried. The precipitate was powdered well andredried at room temperature /0.1 mm/16 hr to afford 1.18 g (91%) of (I)as an orange powder, mp decomposes slowly from 216°. IR (Nujol) 3.00,3.10 μm (OH,NH), 5.95 C═N), 6.15, 6.29 (C═O, chelated quinone), UV-Vismax (CH₃ OH) 233 nm (ε = 88,500), 250 sh (55,900), 288 (14,700), 477(22,000), 495 (21,100), 531 sh (11,800). XL-100 NMR DMSOd₆ (˜ 55°)8.0-14.0 δ (v. broad, OH,N.sup.⊕ H₃), 9.50 (bs, 2, NHCO), 7.81 (m, 4,H-1,3), 7.56 (m, 2, H-2), 5.32 (bs, 2, H-1'), 5.10 (s, 4, OH-4', 9),4.90 (bs, 2, H-7), 4.12 ("d", 2, J=6Hz, H-5'), 3.96 (s, 6, OCH₃), 3.64(bs, 2, H-4'), 3.38 (m, 2, H-3'), 2.98 (m, 4, H-10), 2.37 (bs, 4,(CH₂)₂), ˜ 2.37 (bs, 2, H-8B), 1.90 (s, 6, H-14), ˜ 1.85 (m, 2, H-8A),1.80 (m, 4, H-2'), 1.18 (d, 6, J=6Hz, CH₃ -5'). [α]_(D) ²¹° = +247° (c,0.048, EtOH. Tlc on SiHF CHCl₃ /CH₃ OH/H₂ O (20/10/1 ), (XII) R_(f)0.40, (I) R_(f) 0.04. Paper chromatography on Whatman No. 1 Paper: inn-BuOH/AcOH/H₂ O (5/2/3) (XII) R_(f) 0.73, (I) R_(f) 0.62; inn-PrOH/EtOAc/H₂ O (7/1/2) (XII) R_(f) 0.67, (I) 0.07.

    ______________________________________                                        Anal. Calcd. for C.sub.58 H.sub.64 N.sub.6 O.sub.20 ·2HCl.multido    t.3H.sub.2 O                                                                            C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    53.91    5.62     5.49   6.51                                     Found:      53.81    5.32     5.53   6.61                                     ______________________________________                                    

EXAMPLE 2 Bis(daunomycin) Glutarylhydrazone Dihydrochloride (II)

A mixture of 1.13 g (2.0 mmol) of daunomycin hydrochloride and 0.160 g(1.0 mmol) of glutaric dihydrazide in 120 ml of methanol was stirred atroom temperature in the dark for 3 days. The reaction mixture wasconcentrated in vacuo to about 50 ml and then stirred at roomtemperature for two additional days. To the stirred solution was added125 ml of ether dropwise; the resulting fine precipitate was collected,washed with ether and dried. The precipitate was powdered well andredried at room temperature /0.1 mm/16 hr to afford 1.20 g (93%) of (II)as an orange powder, mp 206°-220° dec. IR (Nujol) 2.98, 3.08 μm (OH,NH),6.00 (C═N), 6.18, 6.31 (C═O, chelated quinone). UV-Vis max (CH₃ OH) 233nm (ε = 81,600), 250 sh (51,200), 290 (14,700), 484-492 plateau(19,800), 499 (20,100) 537 sh (11,600). XL-100 NMR DMSOd₆ (˜ 50°)9.0-12.0 δ (v. broad, OH,N.sup.⊕ H₃), 9.89 (bs, 2, NHCO), 7.84 (m, 4,H-1,3), 7.60 (m, 2, H-2), 5.23 (bs, 2, H-1'), 4.95 (s, 2, OH), 4.87 (bs,2, H-7), 4.12 (m, 2, H-5'), 3.98 (s, 6, OCH₃), 3.63 (bs, 2, H-4'), ˜ 3.3(H-3', hidden by H₂ O peak), ˜ 3.1 (H-10B, hidden by H₂ O peak), 2.91(d, 2, J=19Hz, H-10A), 2.23 (m, 8, H-8, COCH₂ CH₂ CH₂ CO), 1.94 (s, 6,H-14) ˜ 1.94 (m, 2, COCH₂ CH₂ CH₂ CO), 1.77 (m, 4, H-2'), 1.19 (d, 6,J=7Hz, CH₃ -5'). [α]_(D) ^(<)° = + 166° (c, 0.052, EtOH). Tlc on SiHFCHCl₃ /CH₃ OH/H₂ O (20/10/1 ), (XII) R_(f) 0.40 (II) R_(f) 0.07. Paperchromatography on Whatman No. 1 Paper: in n-BuOH/AcOH/H₂ O (5/2/3) (XII)R_(f) 0.73, (II) R_(f) 0.66; in n-PrOH/EtOAc/H₂ O (7/1/2) (XII) R_(f)0.67, (II) R_(f) 0.15.

    ______________________________________                                        Anal. Calcd. for C.sub.59 H.sub.66 N.sub.6 O.sub.20 ·2HCl.multido    t.2 1/2 H.sub.2 O                                                                       C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    54.63    5.67     5.47   6.48                                     Found:      54.67    5.45     5.40   6.65                                     ______________________________________                                    

EXAMPLE 3 Bis(daunomycin) Adipylhydrazone Dihydrochloride (III)

A mixture of 0.316 g (0.56 mmol) of daunomycin hydrochloride and 0.049 g(0.28 mmol) of adipic acid dihydrazide in 25 ml of methanol was stirredat room temperature in the dark. After 5 days, the reaction mixture wasconcentrated to 13 ml and then 15 ml of ether was added dropwise. Theresulting precipitate was collected, washed with 3-10 ml portions ofether and dried at room temperature /0.1 mm/15 hr to yield thebis-hydrazone dihydrochloride (III), 0.333 g (91%), mp - decomposesgradually 235°-250°. IR (Nujol) 2.82, 3.11 μm (OH), 5.98 C═O,hydrazone), 6.19, 6.31 (C═O, chelated quinone), UV-Vis max (CH₃ OH) 233nm (ε = 83,100), 250 sh (51,100), 289 (14,900), 479-488 plateau(20,300), 489-498 plateau (20,200), 535 sh (11,300). XL-100 NMR DMSOd₆(˜ 55°) 8-13 δ (v. broad, OH, N.sup.⊕ H₃), 9.96 (s, 2, CONH), 7.80 (m,4, H-1,3), 7.56 (m, 2, H-2), 5.35 (bs, 2, OH), 5.25 (bs, 2, H-1'), 5.04(bs, 2, OH), 4.74 (m, 2, H-7), 4.12 (m, 2, H-5'), 3.97 (s, 6, OCH₃),3.64 (bs, 2, H-4'), 3.40 (m, 2, H-3'), 3.22 (bs, H₂ O), 2.98 (bs, 4,H-10), 2.30 (m, 8, H-8, COCH₂ (CH₂)₂ CH₂ CO), 1.95 (s, 6, H-14), 1.80(bs, 4, H-2'), 1.49 (bs, 4, COCH₂ (CH₂)₂ CH₂ CO), 1.18 (d, 6, J=6Hz, CH₃-5'). [α]_(D) ²² ° = +247° (c, 0.047, EtOH). Tlc on SiGF in CHCl₃ /CH₃OH/2.0 N AcOH (10/5/1) (XII) R_(f) 0.56, (III) R_(f) 0.30. Paperchromatography in n-PrOH/H₂ O (4/1) (XII) R_(f) 0.63, (III) R_(f) 0.30;in n-BuOH/AcOH/H₂ O (5/2/3) (XII) R_(f) 0.75, (III) R_(f) 0.69.

    ______________________________________                                        Anal. Calcd. for C.sub.60 H.sub.68 N.sub.6 O.sub.20 ·2HCl.multido    t.2 1/2 H.sub.2 O                                                                       C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    54.96    5.77     5.41   6.41                                     Found:      55.02    5.48     5.19   6.30                                     ______________________________________                                    

EXAMPLE 4 Bis(daunomycin) Suberylhydrazone Dihydrochloride (IV)

A mixture of 0.406 g (0.72 mmol) of daunomycin hydrochloride and 0.073 g(0.36 mmol) of suberic acid dihydrazide in 70 ml of methanol was stirredat room temperature in the dark for 3 days. After concentrating to 35ml, the reaction mixture was stirred at room temperature for anadditional 8 days. To the stirred solution was then added 35 ml of etherdropwise; the resulting precipitate was collected, washed with 3-10 mlportions of ether and dried at room temperature /0.1 mm/18 hr to afford0.407 g (85%) of the bis-hydrazone dihydrochloride (IV), mp 229°-232°dec. IR (Nujol) 2.82, 3.02 μm (OH), 5.99 (C═O, hydrazone), 6.20, 6.31(C═O, chelated quinone). UV-Vis max (CH₃ OH) 233 nm (ε = 85,300), 250 sh(53,500), 289 (15,300), 484-492 plateau (20,500), 498 (20,800), 535(12,200). XL-100NMR DMSOd₆ (˜ 55°) 8-12 δ (v. broad, N.sup. ⊕ H₃ OH),9.82 (s, 2, CONH), 7.80 (m, 4, H- 1,3), 7.59 (m, 2, H-2), 5.27 (bs, 2,H-1'), 5.17 (bs, 4, OH), 4.78 (bs, 2, H-7), 4.10 (m, 2, H-5'), 3.97 (s,6, OCH₃), 3.64 (bs, 2, H-4'), 3.36 (m, 2, H-3'), 3.21 (d, 2, J=19HzH-10B), 3.20 (bs, H₂ O), 2.90 (d, 2, J=19Hz H-10A), 2.26 (m, 8 COCH₂(CH₂)₄ CH₂ CO,H-8), 1.93 (s, 6, H-14), 1.82 (bs, 4, H-2'), 1.35 (m, 8,COCH₂ (CH₂)₄ CH₂ CO), 1.18 (d, 6, J=6HzCH₃ -5'). [α]_(D) ²¹° = +273° (c0.053, EtOH). Tlc on SiGF in CHCl₃ /CH₃ OH/2.0 N AcOH (10/5/1) (XII)R_(f) 0.57, (IV) R_(f) 0.34. Paper chromatography in n-PrOH/H₂ O (4/1)(XII) R_(f) 0.71, (IV) R_(f) 0.44; in n-BuOH/AcOH/H₂ O (5/2/3) (XII)R_(f) 0.76, (IV) R_(f) 0.68.

    ______________________________________                                        Anal. Calcd. for C.sub.62 H.sub.72 N.sub.6 O.sub.20 ·2HCl.multido    t.2H.sub.2 O                                                                            C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    55.98    5.91     5.33   6.32                                     Found:      55.96    5.75     5.28   6.27                                     ______________________________________                                    

EXAMPLE 5 Bis(Daunomycin) Sebacylhydrazone Dihydrochloride (V)

A mixture of 0.406 g (0.72 mmol) of daunomycin hydrochloride and 0.083 g(0.36 mmol) of sebacic acid dihydrazide in 165 ml of methanol wasstirred at room temperature in the dark. At intervals, the reactionmixture was concentrated in the following manner: after 4 days to 70 ml,after 9 days to 35 ml, after 11 days to 21 ml, and after 28 days to 14ml. After 35 days, an additional 0.008 g (0.036 mmol) of sebacic aciddihydrazide was added to the reaction mixture. After 42 days, thereaction mixture was diluted with 14 ml of ether added dropwise. Theresulting precipitate was collected, washed with 5-10 ml portions ofether and dried at room temperature/0.1 mm/15 hr to give 0.419 g (87%)of the bis-hydrazone dihydrochloride (V), mp 211°-215° dec. IR (Nujol)2.82, 3.14 μm (OH), 6.00 (C═ O, hydrazone), 6.18, 6.30 (C═O, chelatedquinone). UV-Vis max (CH₃ OH) 234 nm (ε = 82,600), 250 sh (53,700), 288(14,900), 481 (20,600), 488-497 plateau (20,400), 533 sh (11,900).XL-100 NMR DMSOd₆ (˜ 50°) 8-11 δ (v. broad, N.sup.⊕ H₃, OH), 9.86 (bs,2, CONH), 7.79 (m, 4, H-1,3), 7.58 (m, 2, H-2), 5.28 (bs, 4, H-1', OH),4.76 (m, 2, H-7), 4.07 (m, 2, H-5'), 3.95 (s, 6, OCH₃), 3.62 (bs, 2,H-4'), 3.37 (m, 2, H-3'), 3.20 (bs, 2, H-10B), 2.82 (d, 2, J=19Hz,H-10A), 2.21 (m, 8, H-8, COCH₂ (CH₂)₆ CH₂ CO), 1.92 (s, 6, H-14), 1.82(m, 4, H-2'), 1.19 (d, 6, J=6Hz CH₃ - 5'), 1.00 (bs, 12, COCH₂ (CH₂)₆CH₂ CO). [α]_(D) = +260° (c, 0.049, EtOH. Tlc on SiGF in CHCl₃ /CH₃OH/2.0 N AcOH (10/5/1) (XII) R_(f) 0.52, (V) R_(f) 0.31. Paperchromatography in n-PrOH/H₂ O (4/1) (XII) R_(f) 0.71, (V) R_(f) 0.56; inn-BuOH/AcOH/H₂ O (5/2/3), (XII) R_(f) 0.76, (V) R_(f) 0.59.

    ______________________________________                                        Anal. Calcd. for C.sub.64 H.sub.76 N.sub.6 O.sub.20 ·2HCl.multido    t.H.sub.2 O                                                                             C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    57.35    6.02     5.29   6.27                                     Found:      57.30    5.90     4.86   6.31                                     ______________________________________                                    

EXAMPLE 6 Bis(N,N-dimethyldaunomycin) Succinylhydrazone Dihydrochloride(VI)

A mixture of 0.374 g (0.6 mmol) of N,N-dimethyldaunomycin hydrochloride1 3/4 hydrate and 0.0438 g (0.3 mmol) of succinic acid dihydrazide in 36ml of methanol was stirred at room temperature in the dark for 2 days.After concentrating to 12 ml, the reaction mixture was stirred at roomtemperature for 5 additional days and then was diluted with 18 ml ofether added dropwise. The resulting precipitate was collected, washedwith 3-10 ml portions of ether and dried at room temperature/0.1 mm/16hr to yield 0.367 g (91%) of the bis-hydrazone (VI), mp 194°-196°. IR(Nujol) 2.98 μm (OH), 3.70 (HN.sup.⊕ (CH₃)₂), 5.98 (C═O, hydrazone),6.18, 6.31 (C═O, chelated quinone). UV-Vis max (CH₃ OH) 233 nm (ε =89,900), 250 sh (56,700), 288 (15,400), 478 (22,200), 494 sh (21,200),531 sh (11,800). XL-100 NMR DMSOd₆ (˜ 55°) 13.99 δ (s, 2, OH-6), 13.20(s, 2, OH-11), 9.90 (bs, 2. HN.sup.⊕ (CH₃)₂), 9.74 (bs, 2, CONH), 7.79(m, 4, H-1,3), 7.57 (m, 2, H-2), 5.57 (d, 2, J=6Hz, OH-4'), 5.41 (s, 2,H-1'), 5.15 (s, 2, OH-9), 4.91 (m, 2, H-7), 4.05 (m, 2, H-5'), 3.96 (s,6, OCH₃), ˜ 3.9 (m, 2, H-3'), 3.40 (m, 2, H-4'), 3.21 (s, H₂ O), 3.07(bs, 2, H-10B), 2.87 (bs, 2, H-10A), 2.75 (s, 12, N.sup.⊕ (CH₃)₂), 2.30(m, 8, H-8, COCH₂ CH₂ CO), 2.00 (m, 4, H-2'), 1.87 (s, 6, H- 14), 1.19(d, 6, J=6Hz, CH₃ -5'). [α]_(D) = +198° (c 0.051, EtOH). Tlc on SiGF inCHCl₃ /CH₃ OH/2.0 N AcOH (10/5/1) starting material R_(f) 0.48, (VI)R_(f) 0.28. Paper chromatography in n-PrOH/H₂ O (4/1) starting materialR_(f) 0.78, (VI) R_(f) 0.31; in n-BuOH/AcOH/H₂ O (5/2/3) startingmaterial R_(f) 0.75, (VI) R_(f) 0.66.

    ______________________________________                                        Anal. Calcd. for C.sub.62 H.sub.72 N.sub.6 O.sub.20 ·2HCl.multido    t.3H.sub.2 O                                                                            C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    55.23    5.98     5.26   6.23                                     Found:      55.06    5.74     4.87   6.27                                     ______________________________________                                    

EXAMPLE 7 Bis(adriamycin) Succinylhydrazone Dihydrochloride (VII)

A solution of 1.16 g (2.0 mmol) of adriamycin hydrochloride (XI) and0.146 g (1.0 mmol) of succinic acid dihydrazide in 400 ml of methanolwas stirred at room temperature in the dark. The reaction mixture wasconcentrated at intervals in the following manner: after 3 days to 200ml, after 6 days to 100 ml, after 10 days to 50 ml, and after 14 days to25 ml. After 22 days, an additional 0.015 g (0.1 mmol) of succinic aciddihydrazide was added to the reaction mixture. After 44 days, thereaction mixture was slowly diluted with 25 ml of ether; the resultingprecipitate was collected, washed with ether and dried at roomtemperature/0.1 mm/15 hr to afford 1.22 g of (VII) containing 5-7%adriamycin hydrochloride (XI). A 0.608 g sample of crude (VII) in 10 mlof methanol was applied on a 2.5 × 90 cm (˜ 440 ml) column of SephadexLH-20 (packed and washed with methanol). The column was eluted withmethanol and 5.0 ml fractions were collected. Fractions No. 41-54 werecombined and evaporated to yield 0.536 g of (VII). A second 0.607 gsample of crude (VII) was purified in a similar manner. The combinedsample (0.961 g) of purified (VII) was dissolved in 20 ml of methanoland the solution was stirred and diluted with 20 ml of ether addeddropwise. The resulting precipitate was collected, washed with 3-10 mlportions of ether and dried at room temperature/0.1 mm/20 hr to give0.930 g (68%) of the bis-hydrazone dihydrochloride (VII), mp - graduallydecomposes from 210°. IR (Nujol) 3.05 μm (OH), 6.00 (C═O, hydrazone),6.18, 6.32 (C═O, chelated quinone). UV-Vis max (CH₃ OH) 233 nm (ε =86,000), 246 sh (59,400), 288 (15,800), 478 (21,900), 489 sh (21,200),530 sh (11,700). XL-100 NMR DMSOd₆ (˜ 55°), 8-12 δ (v. broad N.sup.⊕ H₃,OH), 10.23 (bs, 2, CONH), 7.80 (m, 4, H-1,3), 7.55 (m, 2, H-2), 5.75(bs, 2, OH), 5.31 (bs, 2, H-1'), ˜ 5.3 (bs, 2, OH), 5.17 (bs, 2, OH),4.91 (m, 2, H-7), 4.43 (bs, 4, H-14), 4.09 (m, 2, H-5'), 3.96 (s, 6,OCH₃), 3.63 (bs, 2, H-4'), 3.40 (m, 2, H-3'), 3.21 (s, H₂ O), 3.05 (d,2, J=19Hz, H-10B), 2.94 (d, 2, J=19Hz, H-10A), 2.31 (m, 4, H-8), 1.83(m, 4, H-2'), 1.19 (d, 6, J=6Hz, CH₃ -5'). [α]_(D) = +232° (c 0.05,EtOH). Tlc on SiGF in CHCl₃ /CH₃ OH/H₂ O (20/10/1), (XI) R_(f) 0.32,(VII) R_(f) 0.03; in CHCl₃ /CH₃ OH/2.0 N AcOH (10/5/1) (XI) R_(f) 0.43,(VII) R_(f) 0.17. Paper chromatography in n-PrOH/H₂ O (4/1) (XI) R_(f)0.47, (VII) R_(f) 0.05; in n-BuOH/AcOH/H₂ O (5/2/3) (XI) R_(f) 0.62,(VII) R_(f) 0.56.

    ______________________________________                                        Anal. Calcd. for C.sub.58 H.sub.64 N.sub.6 O.sub.22 ·2HCl.multido    t.5H.sub.2 O                                                                            C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    51.22    5.63     5.21   6.18                                     Found:      51.21    5.27     5.13   6.36                                     ______________________________________                                    

EXAMPLE 8 Bis(daunomycin) Malonylhydrazone Dihydrochloride (VIII)

A solution of 0.339 g (0.6 mmol) of daunomycin hydrochloride and 0.0396g (0.3 mmol) of malonyl dihydrazide in 24 ml of methanol was stirred inthe dark at room temperature. After 14 days, the reaction mixture wasconcentrated to 12 ml and then 25 ml of ether were added dropwise. Theresulting precipitate was collected, washed with 5--5 ml portions ofether and dried at room temperature /0.1 mm/16 hr to afford 0.351 g(93%) of the malonylhydrazone (VIII), mp dec. 225°-240°. IR (Nujol) 2.92μm (OH), 5.92 (C═N), 6.17, 6.31 (C═O, chelated quinone). UV-Vis max (CH₃OH) 234 nm (ε 75,200), 250 sh 848,400), 287 (12,900), 483 (18,400),493-499 plateau (17,900), 535 sh (10,300). XL-100 NMR DMSOd₆ (˜ 55°)8-12 δ (v. board, OH, N.sup.⊕ H₃), 10.15 (s, 2, CONH), 7.72 (m, 4,H-1,3), 7.47 (m, 2, H-2), 5.29 (bs, 2, OH), 5.21 (bs, 2, H-1'), 4.87(bs, 2, OH), 4.75 (bs, 2, H-7), 4.10 (m, 2, H-5'), 3.95 (s, 6, OCH₃),3.61 (bs, 4, COCH₂ CO, H-4'), 3.37 (m, 2, H-3'), 3.23 (bs, H₂ O), 2.83(bs, 4, H-10), 2.19 (m, 4, H-8), 1.92 (s, 6, H-14), 1.77 (m, 4, H-2'),1.13 (d, 6, J=6Hz, CH₃ -5'). [α]_(D) ²¹° = +230° (c 0.053, EtOH). Tlc onSiGF in CHCl₃ /CH₃ OH. 2 N AcOH (10/5/1/) (XII) R_(f) 0.56, (VIII) R_(f)0.17. Paper chromatography in n-PrOH/H₂ O (4:1): (XII) R_(f) 0.56,(VIII) R_(f) 0.09; in n-BuOH/AcOH/H₂ O (5/2/3): (XII) R_(f) 0.79, (VIII)R_(f) 0.71.

    ______________________________________                                        Anal. Calcd. for C.sub.57 H.sub.62 N.sub.6 O.sub.20 ·2HCl.multido    t.1 1/2 H.sub.2 O                                                                       C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    54.72    5.40     5.67   6.72                                     Found:      54.84    5.59     5.43   6.71                                     ______________________________________                                    

EXAMPLE 9 Bis-(Daunomycin) Oxalylhydrazone Dihydrochloride (IX)

A suspension of 0.564 g (1.0 mmol) of daunomycin hydrochloride and 0.059g (0.5 mmol) of oxalyl dihydrazide in 200 ml of methanol was stirred atroom temperature in the dark. After 15 days, the homogenous reactionmixture was concentrated to 100 ml and stirred at room temperature foran additional 40 days. The reaction mixture was concentrated to 50 mland then 75 ml of absolute ethanol was added dropwise. The resultingfine gelatinous precipitate was collected by centrifugation (3000 X g);the precipitate was transferred to a fritted disc funnel and washed with2--2 ml portions of absolute ethanol and with 5 ml of ether. Afterdrying at room temperature, the precipitate was powdered well andredried at room temperature /0.1 mm/17 hr to afford 0.459 g (73%) of thebis-hydrazone dihydrochloride (IX), mp gradually dec. 220°-240°. IR(Nujol) 2.90, 3.04 μm (OH), 5.99 (C═O, hydrazone), 6.16, 6.29 (C═ O,chelated quinone). UV-Vis max (CH₃ OH) 233 nm (ε = 83,200), 249(57,700), 284 sh (17,400), 478 (22,700), 496 (22,000), 531 sh (12,000).XL-100 NMR DMSOd₆ (˜ 50°) 6-12 δ (v. broad, N.sup.⊕ H₃, OH), 10.50 (bs,2, CONH), 7.82 (m, 4, H-1,3), 7.63 (m, 2, H-2), 5.29 (bs, 4, H-1', OH),4.89 (m, 2, H-7), 4.13 (m, 2, H-5'), 3.98 (s, 6, OCH₃), 3.61 (bs, 2,H-4'), 3.34 (m, 4, H-10B, H-3'), 3.18 (s, H₂ O), 2.89 (d, 2, J=19Hz,H-10A), 2.20 (m, 4, H-8), 1.94 (s, 6, H-14), 1.80 (m, 4, H-2'), 1.17("bs," 6, CH₃ -5'). [α]_(D) ²¹° = +261° (c 0.046, CH₃ OH). Tlc on SiGFCHCl₃ /CH₃ OH/2 N AcOH (10/5/1) (XII) R_(f) 0.49, (IX) R_(f) 0.20. Paperchromatography in n-PrOH/H₂ O (4/1): (XII) R_(f) 0.66 (IX) R_(f) 0.02;in n-BuOH/AcOH/H₂ O (5/2/3) (XII) R_(f) 0.73, (IX) R_(f) 0.68.

    ______________________________________                                        Anal. Calcd. for C.sub.56 H.sub.60 N.sub.6 O.sub.20 ·2HCl.multido    t.2 1/2 H.sub.2 O (1255.08)                                                             C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    53.59    5.38     5.65   6.70                                     Found:      53.56    5.51     5.45   6.44                                     ______________________________________                                    

EXAMPLE 10 Bis-(daunomycin) Pimelylhydrazone Dihydrochloride (X)

A mixture of 0.452 g (0.8 mmol) of daunomycin hydrochloride and 0.0753 g(0.4 mmol) of pimelyl dihydrazide in 50 ml of methanol was stirred atroom temperature in the dark for 7 days during which time thebis-hydrazone (X) precipitated. The precipitate was collected, washedwith 5-1 ml portions of methanol and with 3-5 ml portions of ether anddried at room temperature. The precipitate was powdered well and redriedat room temperature /0.1 mm/17 hr to give 0.373 g (71%) of thebis-hydrazone (X), mp 239°-242° dec. IR (Nujol) 2.80, 3.08 μm (OH), 6.00(C═O, hydrazone), 6.16, 6.29 (C═O, chelated quinone). UV-Vis max (CH₃OH) 233 nm (ε 85,800), 250 sh (54,200), 290 (16,200), 486 (21,300), 499(21,400), 536 sh (12,300). XL-100 NMR DMSOd₆ (˜ 50°) 8-12 δ (v. broad,N.sup.⊕ H₃, OH), 9.81 (s, 2, CONH), 7.81 (m, 4, H-1,3), 7.58 (m, 2,H-2), 5.25 (bs, 4, H- 1', OH), 5.15 (bs, 2, OH), 4.83 (m, 2, H-7), 4.10(m, 2, H-5'), 3.96 (s, 6, OCH₃), 3.64 (bs, 2, H4'), 3.24 ("m," 4, H-3',H-10B), 3.19 (bs, H₂ O), 2.81 ("d," 2, J=19Hz, H-10A), 2.25 (m, 8, H-8,COCH₂ (CH₂)₃ CH₂ CO) 1.94 (s, 6, H-14), 1.81 (m, 4, H-2'), 1.32 (m, 6,COCH₂ (CH₂)₃ CH₂ CO), 1.19 (d, 6, J=6Hz, CH₃ -5'). [α]_(D) ²¹° = +239°(c0.054, CH₃ OH). Tlc on SiGF CHCl₃ /CH₃ OH/2 N AcOH (10/5/1 ) (XII) R_(f)0.50, (X) R_(f) 0.23. Paper chromatography in n-PrOH/H₂ O (4/1) (XII)R_(f) 0.66 (X) R_(f) 0.27; in n-BuOH/AcOH/H₂ O (5/2/3) (XII) R_(f) 0.73,(X) R_(f) 0.68.

    ______________________________________                                        Anal. Calcd. for C.sub.61 H.sub.70 N.sub.6 O.sub.20 ·2HCl.multido    t.1 1/2 H.sub.2 O (1307.21)                                                             C      H        Cl.sup.θ                                                                         N                                          ______________________________________                                                    56.05    5.78     5.42   6.43                                     Found:      55.89    5.47     5.36   6.47                                     ______________________________________                                    

BIOLOGICAL TESTS

Biological testing data for compounds of this invention as the HClsalts, as well as for adriamycin and daunomycin, are presented in thetable given below. The compounds were tested first in cultured lymphoidleukemia L1210 cells for inhibition of nucleic acid synthesis bypreviously reported procedures. The analogs were then tested againstlymphocytic leukemia P388 implanted in mice, under the auspices of theNCI and according to its protocols which use the increased survival timeof treated mice compared to controls as the measure of antitumorefficacy.

    __________________________________________________________________________    Bioassay Data on Bis-hydrazones of                                            Daunomycin and Adriamycin                                                                Vs Cultured L1210 Leukemia Cells.sup.b                                                          Vs P388 Leukemia in Mice.sup.c                              Inhibition of Synthesis                                                                         Optimum                                                                              Antitumor                                                                           Optimum Dose                               NSC.sup.a                                                                         of DNA       of RNA                                                                             Dose, qd 1-9                                                                         Efficacy                                                                            q4d5,9,13                                                                             Antitumor Efficacy          Compound                                                                             No. ED.sub.50, μM                                                                           Ed.sub.50, μM                                                                   mg/kg  T/C, %                                                                              mg/kg   T/C,%                       __________________________________________________________________________    I      266210                                                                            13           3.4  6.25   283.sup.d                                                                           20      170                                                      12.5   307.sup.e                                 II     266211                                                                            4.5          3.6  1.56   188                                       no data--                                                                                                  3.12   149                                       III    273432                                                                            8.9          3.6  6.25   281,289                                                                             37.5    196                         IV     276747                                                                            4.9          2.4  8      245   25      178                         V      276748                                                                            10           1.0  12.5   221   50      163                         VI     274885                                                                            19           3.0  4      178   25      158                         VII    273433                                                                            13.3         3.1  6.25   309   18.8    160                         VIII   279510                                                                            14.1         7.4  6.25   169   (inactive)                          IX     285695                                                                            4.3          2.3               25      160                         X      285696                                                                            2.0          1.3               25      160                         Adriamycin                                                                           123127                                                                            1.5          0.67 1.0    195   8       157                         Daunomycin                                                                           82151                                                                             0.49         0.39 0.5    166   8       134                         __________________________________________________________________________     .sup.a Accession number of the National Cancer Institute.                     .sup.b Assay described in G. Tong, W. W. Lee, D. R. Black and D. W. Henry     J. Med. Chem., 19 395 (1976).                                                 .sup.c Ip P388 murine leukemia treated ip on QD1-9 and Q4D 5, 9, 13           schedules according to standard NCI protocols. Assay described in R. I.       Geran, N. H. Greenberg, M. M. MacDonald, A. M. Schumacker and B. J.           Abbott, Cancer Chemother. Rep., Part 3, 3 (No. 2), 9 (1972), Protocol         1,200. T/C = ratio of survival time of treated mice to that of untreated      controls times 100. Untreated controls survive about 9 days.                  .sup.d Three out of six treated mice survived until sacrifice at 30 days.     .sup.e Two out of six treated mice survived until sacrifice at 45 days.  

We claim:
 1. Compounds having the structure ##STR3## wherein n is aninteger having a value of from 0 through 8 and the radicals indicated byR and R' each represent hydrogen except that in those instances in whichn has a value of 2, the R groups, if not hydrogen, may be methylradicals (with R' remaining as hydrogen), and the R' groups, if nothydrogen, may be hydroxy radicals (with R remaining as hydrogen),together with their pharmaceutically acceptable acid addition salts. 2.The compound of claim 1 in which is bis(daunomycin) succinylhydrazonetogether with its pharmaceutically acceptable acid addition salts. 3.The compound of claim 1 which is bis(daunomycin) glutarylhydrazonetogether with its pharmaceutically acceptable acid addition salts. 4.The compound of claim 1 which is bis(daunomycin) adipylhydrazonetogether with its pharmaceutically acceptable acid addition salts. 5.The compound of claim 1 which is bis(daunomycin) suberylhydrazonetogether with its pharmaceutically acceptable acid addition salts. 6.The compound of claim 1 which is bis(daunomycin) sebacylhydrazonetogether with its pharmaceutically acceptable acid addition salts. 7.The compound of claim 1 which is bis(N,N-dimethyldaunomycin)succinylhydrazone together with its pharmaceutically acceptable acidaddition salts.
 8. The compound of claim 1 which is bis(adriamycin)succinylhydrazone together with its pharmaceutically acceptable acidaddition salts.
 9. The compound of claim 1 which is bis(daunomycin)malonylhydrazone together with its pharmaceutically acceptable acidaddition salts.
 10. The compound of claim 1 which is bis(daunomycin)oxalylhydrazone together with its pharmaceutically acceptable acidaddition salts.
 11. The compound of claim 1 which is bis(daunomycin)pimelylhydrazone together with its pharmaceutically acceptable acidaddition salts.